Yin Xiao-Cheng, Peng Yan-Hui, Xiao Zheng-Xiang
Department of Pediatrics, The First Affilliated Hospital of Nanhua University, Hengyang 421001, Hunan Province, China.
Zhongguo Shi Yan Xue Ye Xue Za Zhi. 2011 Jun;19(3):634-7.
The aim of this study was to investigate the effect of diallyl disulfide (DADS) on the apoptosis of K562 cells and to explore the mechanism of K562 apoptosis induced by DADS. The K562 cells were treated with different concentrations of DADS for 24, 48 and 72 hours. The concentrations of DADS were as follows: 0 (control group), 10, 20, 40 and 80 mg/L. The morphologic changes of leukemia K562 cells treated with DADS were observed by Hoechst33 258 staining. The apoptosis of K562 cells treated with different concentrations of DADS for 24, 48 and 72 hours was analyzed by flow cytometry. The mRNA expression changes of Fas and FasL were detected by reverse transcription-polymerase chain reaction (RT-PCR) after K562 cells were treated with different concentrations of DADS for 48 hours. The results indicated that the characteristics of apoptosis in K562 cells induced by DADS were as follows: reduction of nucleus, chromatin condensation and nuclear membrane rupture. The flow cytometry with PI straining showed that after 24 hours of DADS treatment the apoptosis rate of K562 cells increased from 11.60 ± 0.83% at the concentration of 10 mg/L to 37.94 ± 0.87% at the concentration of 40 mg/L. The apoptosis rate of K562 cells increased from 37.94 ± 0.87% (24 hours) to 47.02 ± 0.66% (72 hours) after treatment with DADS of 10 mg/L increasing to 40 mg/L DADS. The Fas mRNA expression levels of the related apoptotic genes increased after K562 cells were treated with different concentrations of DADS for 48 hours, while FasL mRNA expression decreased significantly after DADS treatment for 48 hours, compared with those in the control group (p < 0.05). It is concluded that DADS can induce the apoptosis of human leukemia K562 cells in a time-and concentration-dependent manners. The activation of Fas/FasL pathway may play an important role in the K562 cell apoptosis induced by DADS, which is associated with increasing Fas gene expression and decreasing FasL gene expression.
本研究旨在探讨二烯丙基二硫醚（DADS）对K562细胞凋亡的影响，并探究DADS诱导K562细胞凋亡的机制。将K562细胞用不同浓度的DADS处理24、48和72小时。DADS的浓度如下：0（对照组）、10、20、40和80mg/L。采用Hoechst33258染色观察经DADS处理的白血病K562细胞的形态变化。用流式细胞术分析不同浓度DADS处理24、48和72小时后K562细胞的凋亡情况。K562细胞用不同浓度DADS处理48小时后，通过逆转录-聚合酶链反应（RT-PCR）检测Fas和FasL的mRNA表达变化。结果表明，DADS诱导K562细胞凋亡的特征如下：细胞核缩小、染色质浓缩和核膜破裂。PI染色的流式细胞术显示，DADS处理24小时后，K562细胞的凋亡率从10mg/L浓度时的11.60±0.83%增加到40mg/L浓度时的37.94±0.87%。用10mg/L至40mg/L的DADS处理后，K562细胞的凋亡率从24小时时的37.94±0.87%增加到72小时时的47.02±0.66%。K562细胞用不同浓度DADS处理48小时后，相关凋亡基因的Fas mRNA表达水平升高，而DADS处理48小时后，FasL mRNA表达与对照组相比显著降低（p<0.05）。结论是，DADS能以时间和浓度依赖性方式诱导人白血病K562细胞凋亡。Fas/FasL途径的激活可能在DADS诱导的K562细胞凋亡中起重要作用，这与Fas基因表达增加和FasL基因表达降低有关。
